28 August 2006
Connective Tissues Part 1
Dr. Robert Crissman, Ph.D.



Definition


  • All connective tissue is composed of cells, extracellular fibers and ground substances
  • Group of interrelated tissues generally classified as “connective tissue”
  • Connects all the other cells of different tissues types both structurally and metabolically
  • Provides and maintains the form of body
  • Matrix that connects and binds cells tissues, and organs together
  • All connective tissues are derived from a common origin of embryonic mesenchyme

Classification of Connective Tissues

  • Embryonic
    • Mesenchyme – Embryo and Fetus
    • Mucoid Connective Tissue – Umbilical Cord
  • Connective Tissue “Proper”
    • Areolar Connective Tissue – Loose packing (few extracellular fibers) and surrounds blood vessels (BV)
      • All blood vessels are surrounded by connective tissues
    • Dense Connective Tissues – lots of extracellular fibers
      • Dense irregular connective tissue – Dermis, organ capsules, and periosteum
        • Fibers in all sorts of directs
      • Dense regular connective tissue – tendons, ligaments
        • Fibers all run parallel in sheets
      • Elastic connective tissue – ligamentum nuchae and fava
        • Instead of lots of collagen fibers, you have lots of elastin fibers
        • A type of regular connective tissue
    • Reticular tissue – lymphatic tissue and bone marrow
        • Can be classified as loose connective tissue
    • Adipose tissue – subcutaneous and omentum
  • Special Connective Tissue
    • Cartilage – trachea, pina, nose, joint cartilage
    • Bone -- skeleton
    • Blood – cardiovascular and hemopoietic tissue

Functions of Connective Tissue


  • Mechanical support
    • Structural support of organs
      • e.g. connective tissue surrounding gland to support gland
      • Connective tissue of any organ is called stroma and supports the parenchyma
      • Parenchyma is the functional part of an organ supported by the stroma
    • Packing material
      • “Mother nature’s mortar”
      • Fills in all that’s between
    • Rigid structural support
  • Mechanical Protection
    • Bony and cushioning
      • e.g. thoracic cage, skull, heel of foot
  • Energy storage and temperature regulation
    • Fat – stored energy and insulator
  • Metabolic support
    • e.g. blood vessel must go through stroma to get to parenchyma
      • stroma determine transportability of molecules and waste to parenchyma
  • Transportation of material
  • Protection against infection
    • e.g. immune system does most of its work in connective tissue
      • Inflammation is in connective tissue
  • Repair after injury

Composition of All Connective Tissues


  • Cells
    • Fixed
      • Responsible for synthesis and maintenance of extracellular matrix and particular connective tissue
      • Secretory cells secreting extracellular matrix and ground substances
      • e.g. fibrocytes, mesenchymal cells, reticular cells, adipocytes, osteocytes, and chondrocytes
    • Wandering
      • Transient cells that migrate in and out of extracellular matrix of connective tissue
      • e.g. macrophages, mast cells, plasma cells, leukocytes
    • Associated
      • Cells associated with blood vessels that are always within connective tissue
      • e.g. endothelial cells, smooth muscle cells, and pericytes
      • Relatively acellular with lots of extracellular space between cells filled by extracellular matrix
      • Extracellular matrix includes extracellular fibers and amorphous ground substances
  • Extracellular Fibers
    • Collagen fibers for strength
    • Reticular fibers for delicate support
    • Elastic fibers for stretch and flexibility
  • Amorphous Ground Substances
    • Interstitial fluid (water)
    • Proteoglycans
      • Complex macromolecules consisting of core protein with glycoaminoglycans (GAG) attached to it
    • Glycoproteins

Fixed Cells


Fibrocytes

  • Most common connective tissue proper
  • Vesicular nucleus with nucleolus – hard to identify cytoplasm
    • Probably mostly euchromatin visible on nucleus
  • Secretory cell
  • Fibroblast – “-blast” is still capable of division and capable of adding new matrix around it
  • Fibrocyte – “-cyte” terminally differentiated, incapable of dividing, and maintain existing matrix but can’t add new matrix
  • Can’t tell fibroblast from fibrocyte unless the fibroblast is dividing

Mesenchymal Cell

  • Looks and does the same thing as a fibroblast
    • Identify mesenchymal or fibrocyte by location
  • Only difference is that its location is pluripotential
    • Can differentiate to muscle, bone, cartilage, etc.

Reticular cells

  • Similar morphology to fibrocyte
  • Can identify reticular fibers around them
  • Secretory
    • Glycoproteins
  • Forms cell-covered fiber framework

White adipocytes

  • Diagnostic characteristic – lipid droplet is all one lipid droplet (unilocular)
  • Large cells with signet ring appearance with nucleus pushed to one side
  • Adipose tissue is called subcutaneous fascia
  • Function – lipid storage
    • Hypercellular obesity – increased number of fat cells
      • Worse type to have and thought to be caused by overfeeding during the first year of life
      • Hardest one to treat
    • Hypertrophic obesity – increased size of fat cells
      • Most common condition

Lipid Storage

  • Lipoprotein Lipase
    • Put fatty acids into adipose cell
  • Hormone Sensitive Lipase
    • Breakdown triglycerides to glycerol and fatty acids
    • Takes fatty acids out of adipose cell
  • Leptin
    • Protein produced by adipocyte
    • Targets hypothalamus to decrease food intake and increase energy consumption
  • Insulin
    • Stimulates insulin receptors on fat cells to synthesize and store lipids
  • Epinepherine and norepinepherine
    • Stimulate fat cells to break down lipids and release glycerol and free fatty acids

Brown fat cells

  • Smaller than white fat cell
  • Multilocular – several small lipid lakes in cytoplasm
  • Numerous mitochondria on EM level
  • Function is to produce heat
    • Free fatty acid uncouples oxidation process from ATP production
    • Thermogenin on mitochondrial membrane protein that permits back flow of H+ protons instead of using for ATP formation
    • “Parasite Eve”
    • Found in newborns that is thought to gradually be converted to white fat in the first year
      • Newborn mediastinum, aorta, axilla between scapula
    • Also found in hibernating animals

Wandering cells


Marcophage

  • “Big Eater”
  • Antigen presenting cell
  • Have lots of phagosomes, lysosomes, and phagolysosomes
  • Mono-nuclear phagocyte system
  • Derived from granulocytic-monocyte stem cell
    • Transported as monocytes in blood stream
    • When a monocyte leaves the blood and enters connective cell, it becomes a macrophages
    • Name dependent on location where monocyte leaves blood stream
  • Identify with vital dye ingestion such as Indian ink, trypan blue, carbon black
    • Can’t see cytoplasm
    • Foreign-body giant cell – in an inflammation situation, several macrophages fuse to surround and encapsulate foreign-body

Mast Cell

  • Full of granules
    • contain mediators of inflammatory response such as histamine, heparin, neutral protease, leukotrienes (C4 and D4), Eosinophilic chemotaxtic factor, neutrophilic chemotaxtic factor
  • Associated with small blood vessels
  • Antibody attaches to IgE receptor which interacts with antigen
    • Secondary signal via cAMP and phosophylation is activated that leads to activation of phospholipases and Ca2+ release which triggers release of granules
    • Important because of anaphylactic shock
      • Fluid leaking out of blood and entering connective tissue
      • Resulting swelling and dilation of blood vessels on a large scale
      • Blood pressure drops and patient goes into anaphylactic shock
      • Histamine also constricts smooth muscle in bronchia
  • Also releases cytokines such as TNFγ
  • Paracrine secretion – localized secretion that isn’t distributed throughout the body

Plasma cells

  • Morphologically eccentrically located nucleus (nucleus not in center)
  • Has cart wheel appearance
  • Packed with rough ER (visible on EM level)
    • Produces immunoglobulins (antibodies) and needs lots of rough ER
  • Terminally differentiated from B-lymphocytes
  • If shows up on a blood smear, patient has a massive infection somewhere

Extracellular Fibers


Collagen Fibers

  • Strength
  • Most common protein of the body
    • Lots of collagen fiber diseases
    • 20% of all proteins
    • 30% of dry weight of body
  • High tensile strength, inelastic yet flexible
  • Use as model of fiber organization and formation
  • Collagen can be organized into fibrils which are organized into fibers
  • Fibers are organized into bundles or fascicle
    • Fibers are barely visible on light microscope
    • Fibrils are made of trophocollagen molecules which are made of alpha chains
      • Each alpha chain is 1,000 amino acids containing lots of glycine, hydroxyproline, and hydroxylysine
        • Glycine is used as a diagnostic test for collagen wasting diseases
        • H+ bonding between hydroxyproline holds alpha chains together
        • Covalent bonds between lydroxylysine of adjacent tropocollagen molecules hold fibrils together
    • Gap is offset by a quarter of their length, hence “quarter stagger theory”

Types of Collagen

  • Type I
    • Strength
    • Most common type
    • Dermis, tendon, organ capsules, bone, dentin
  • Type II
    • Resists pressure
    • Hyaline and elastic cartilage
  • Type III
    • Delicate support meshwork
    • Lymphatic system, spleen, liver, lung, cardiovascular system
    • Make reticular fibers
  • Type IV
    • Attachment, filtration
    • Basal lamina

Collagen Formation

  • Transcription in nucleus
  • Translation to form alpha chains in Rough ER
  • Hydroxylate and glycosylated in Rough ER
  • Formation of procollagen in Rough ER
    • Has a tag at the end of them to prevent procollagen from self-aggregating inside the cell
  • Packaged and secreted via golgi through merocrine secretion
  • Cleavage to form tropocollagen by procollagen peptidase
    • Tropocollagen self-aggregates and lines up quarter-staggared to form fibrils
  • Self-assembly to form collagen fiber via lysyl oxidase

Defects in Collagen Formation

  • Scurvy
    • Vitamin C needed to hydroxylate proline
    • No hydroxylation means no hydrogen bonds and instabilitly
    • Collagen literally melts in body temperature
    • Capillary collagen collapse and cause little bleeds
    • Teeth held in sockets by ligaments; collapse of collagen and teeth falls out
    • Bruise very easily
  • Ehler-Danlos VII
    • Mutation in procollagen peptidase
    • No cleavage to form tropocollagen
    • Weaken stability
    • Tend to dislocate very easily and be super-flexible
  • Collagen wasting disease
    • Mutations in collagenase increase collagen turn over rate
    • High amounts of glycine in urine
      • Normally, collagen is turned over at a certain rate and needs to be replaced

Reticular Fibers

  • Branched network
  • Diagnostic: intersect at right angles
  • Argyophilic – silver loving
    • Taken up by polysaccharides
  • For delicate support
    • Contains lots of wandering cells
    • Needs space for wandering cell but support to keep spaces open
  • EM level shows that each fiber contains bundles of type III collagen fibrils surrounded by a sugar coat
    • Cross striations visible from type III collagen fibrils
  • Located in areolar connective tissue, liver, reticular, lymphatic and hemopoietic tissue
  • Ehlers-Danlos Type IV
    • Deficiency in Type III Collagen
    • Apparent in aorta which form aneurisms with form easily

Elastic fibers

  • Continuous network of “Y” branching fibers
  • Intersect at acute angles
    • “Chicken-wire” networks
  • Function is to stretch, up to 150x its original shape, reversibly
  • Location: blood vessels, lungs (forcing air back out of alveoli)
  • Secreted like collagen – have trophoelastin and proelastin precursors, etc.
  • EM morphology:
    • Amorphous elastin
      • Desmosine
      • Isodesmosine
    • Microfibrils

Disease of Elastin fibers

  • Marfan’s Syndome
    • Microfibrils not produced
      • G.I. problems
      • Tendency to rupture aorta and other blood vessels
  • Emphysema
    • Elastin fibers are damaged and regrow incorrectly
    • No longer able to force air out of alveoli
      • Become barrel cavity

Amorphous Ground Substances


  • Whats left after you remove all cells and fibers
  • No visible structure

Proteoglycans

  • Slippery and viscous
  • Protein core with sulfated glycoaminocylcans (GAGs) attached
    • In 3-D, they look like a bottle brush
  • 6 types of GAG’s with different GAG’s or protein core to get a different proteoglycan
  • Common Proteoglycans:
    • Free
      • Aggrecan – most common
        • In collagen, Aggrecan links to chondroitin sulfate to form aggregates
      • Perlecan – has heprin sulfate
    • Transmembrane
      • Syndecan – has heparin sulfate and chondroitin sulfate
      • Fibroglycan

GAGs

  • Hyaluronic Acid
    • Non-sulfated
    • Long Chain of repeated disaccharides
    • Will swell up and takes up all the space it can
    • Negatively charged and attracts cations like Ca2+ and Na+ which in turn pulls in water
    • Slippery and resilient gel-like consistency that can resists compression
    • Forms physical barrier – bacteria produce hyaluronidases to break down barriers
    • Inhibits cell adhesion and facilitate cell migration
  • Sulfated GAGs
    • Keratin Sulfate in cartilage
    • Heparan Sulfate in basement membranes
    • Condroitin-6-Sulfate in cartilage, skin and bone
    • Condroitin-4-Sulfate in cartilage, skin and bone
    • Dermatan sulfate in dermis

Interstitial Fluid

  • Diffusion of nutrients and waste

Glycoproteins

  • Adhesion molecules